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CRISPR/Cas9-mediated gene knockout is insensitive to target copy number but is dependent on guide RNA potency and Cas9/sgRNA threshold expression level

Overview of attention for article published in Nucleic Acids Research, September 2017
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About this Attention Score

  • In the top 25% of all research outputs scored by Altmetric
  • High Attention Score compared to outputs of the same age (87th percentile)
  • High Attention Score compared to outputs of the same age and source (90th percentile)

Mentioned by

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33 tweeters

Citations

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20 Dimensions

Readers on

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134 Mendeley
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Title
CRISPR/Cas9-mediated gene knockout is insensitive to target copy number but is dependent on guide RNA potency and Cas9/sgRNA threshold expression level
Published in
Nucleic Acids Research, September 2017
DOI 10.1093/nar/gkx843
Pubmed ID
Authors

Garmen Yuen, Fehad J. Khan, Shaojian Gao, Jayne M. Stommel, Eric Batchelor, Xiaolin Wu, Ji Luo

Abstract

CRISPR/Cas9 is a powerful gene editing tool for gene knockout studies and functional genomic screens. Successful implementation of CRISPR often requires Cas9 to elicit efficient target knockout in a population of cells. In this study, we investigated the role of several key factors, including variation in target copy number, inherent potency of sgRNA guides, and expression level of Cas9 and sgRNA, in determining CRISPR knockout efficiency. Using isogenic, clonal cell lines with variable copy numbers of an EGFP transgene, we discovered that CRISPR knockout is relatively insensitive to target copy number, but is highly dependent on the potency of the sgRNA guide sequence. Kinetic analysis revealed that most target mutation occurs between 5 and 10 days following Cas9/sgRNA transduction, while sgRNAs with different potencies differ by their knockout time course and by their terminal-phase knockout efficiency. We showed that prolonged, low level expression of Cas9 and sgRNA often fails to elicit target mutation, particularly if the potency of the sgRNA is also low. Our findings provide new insights into the behavior of CRISPR/Cas9 in mammalian cells that could be used for future improvement of this platform.

Twitter Demographics

The data shown below were collected from the profiles of 33 tweeters who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 134 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 134 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 24 18%
Student > Ph. D. Student 24 18%
Student > Bachelor 24 18%
Student > Master 15 11%
Other 9 7%
Other 17 13%
Unknown 21 16%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 45 34%
Agricultural and Biological Sciences 39 29%
Engineering 7 5%
Medicine and Dentistry 5 4%
Immunology and Microbiology 5 4%
Other 7 5%
Unknown 26 19%

Attention Score in Context

This research output has an Altmetric Attention Score of 16. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 15 November 2017.
All research outputs
#978,983
of 13,255,714 outputs
Outputs from Nucleic Acids Research
#923
of 21,110 outputs
Outputs of similar age
#33,711
of 270,911 outputs
Outputs of similar age from Nucleic Acids Research
#28
of 285 outputs
Altmetric has tracked 13,255,714 research outputs across all sources so far. Compared to these this one has done particularly well and is in the 92nd percentile: it's in the top 10% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 21,110 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 5.2. This one has done particularly well, scoring higher than 95% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 270,911 tracked outputs that were published within six weeks on either side of this one in any source. This one has done well, scoring higher than 87% of its contemporaries.
We're also able to compare this research output to 285 others from the same source and published within six weeks on either side of this one. This one has done particularly well, scoring higher than 90% of its contemporaries.